Generation and characterization of Chd7-iCreERT2-tdTomato mice

IF 2.4 4区 生物学 Q2 DEVELOPMENTAL BIOLOGY genesis Pub Date : 2023-11-22 DOI:10.1002/dvg.23575
Zi'ang Han, Ze Wang, Zhuxi Huang, Weijun Feng
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Abstract

Heterozygous mutation of CHD7 gene causes a severe developmental disorder called CHARGE syndrome. In order to further explore the expression and function of Chd7 in vivo, we generated a Chd7-P2A-iCreERT2-P2A-tdTomato (in short, Chd7-CT-tdT) knockin mouse line using the CRISPR/Cas9 technology. The specificity and efficiency of two knockin genetic elements were validated. The Chd7-CT-tdT reporter gene could accurately reflect both the dynamic expression pattern of endogenous Chd7 during neurodevelopment and cell-type specific expression in the brain and eye. The recombination efficiency of Chd7-CT-tdT in postnatal cerebellum is very high. Moreover, lineage tracing experiment showed that Chd7 is expressed in intestinal stem cells. In summary, the newly constructed Chd7-CT-tdT mouse line provide a useful tool to study the function of Chd7.

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Chd7-iCreERT2-tdTomato小鼠的产生和表征。
CHD7基因的杂合突变会导致严重的发育障碍,称为CHARGE综合征。为了进一步探索Chd7在体内的表达和功能,我们利用CRISPR/Cas9技术构建了Chd7- p2a - icreert2 - p2a - tdtomato(简称Chd7- ct - tdt)敲入小鼠细胞系。验证了两个敲入基因元件的特异性和效率。Chd7- ct - tdt报告基因既能准确反映内源性Chd7在神经发育过程中的动态表达模式,又能准确反映脑、眼细胞类型特异性表达。Chd7-CT-tdT在出生后小脑中的重组效率非常高。此外,谱系追踪实验表明Chd7在肠干细胞中表达。综上所述,新构建的Chd7- ct - tdt小鼠系为研究Chd7的功能提供了有益的工具。
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来源期刊
genesis
genesis 生物-发育生物学
CiteScore
3.60
自引率
0.00%
发文量
40
审稿时长
6-12 weeks
期刊介绍: As of January 2000, Developmental Genetics was renamed and relaunched as genesis: The Journal of Genetics and Development, with a new scope and Editorial Board. The journal focuses on work that addresses the genetics of development and the fundamental mechanisms of embryological processes in animals and plants. With increased awareness of the interplay between genetics and evolutionary change, particularly during developmental processes, we encourage submission of manuscripts from all ecological niches. The expanded numbers of genomes for which sequencing is being completed will facilitate genetic and genomic examination of developmental issues, even if the model system does not fit the “classical genetic” mold. Therefore, we encourage submission of manuscripts from all species. Other areas of particular interest include: 1) the roles of epigenetics, microRNAs and environment on developmental processes; 2) genome-wide studies; 3) novel imaging techniques for the study of gene expression and cellular function; 4) comparative genetics and genomics and 5) animal models of human genetic and developmental disorders. genesis presents reviews, full research articles, short research letters, and state-of-the-art technology reports that promote an understanding of the function of genes and the roles they play in complex developmental processes.
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